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Adenine Deaminase from Various Sources: Heterologous Expression, Activity Screening, Characterization, and Adenine
Junhao Xu1, Peng Zhou2, Yihong Bao1
1School of Life Sciences, Northeast Forestry University, Harbin 150040, China.
Abstract:
Adenine deaminase (ADE) plays a critical role in food purine degradation and in preventing hyperuricemia. Herein, the ADE genes from four different sources were expressed using pMA5, pMA0911, and pP43NMK as the vectors and Bacillus subtilis WB800 as the host. B. subtilis WB800/p43-AK-ADE exhibited the highest ADE activity of 45.41 U/mL. The optimum pH and temperature for AK-ADE were 7.5 and 45 °C, respectively. The enzyme remained stable within pH 6.0-8.5 and temperature 35-45 °C, respectively. The Km and Kcat values were 0.30 mM and 0.28 s-1, respectively. Furthermore, molecular docking and active pocket prediction revealed that AK-ADE binds to adenine with the lowest binding free energy, indicating the strongest affinity. In contrast, ADE from other sources exhibited markedly reduced binding and suboptimal residue positioning. Finally, when AK-ADE was applied to beer wort, it resulted in the removal of 71% of the adenine content, establishing its potential for industrial application.
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